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- Peptide RTSDSRDHTRVDWKR(C), corresponding to amino acid residues 271-285 of rat GluR1 (Accession P19490). Extracellular, N-terminus.
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Western blot analysis of rat (lanes 1 and 3) and mouse (lanes 2 and 4) brain lysates:1,2. Anti-GluR1 (GluA1) (extracellular) Antibody (#AGC-004), (1:200).
3,4. Anti-GluR1 (GluA1) (extracellular) Antibody, preincubated with the control peptide antigen.
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Expression of GluR1 in mouse hippocampusImmunohistochemical staining of mouse hippocampus with Anti-GluR1 (GluA1) (extracellular) Antibody (#AGC-004). GluR1 (green) is present in the stratum oriens (Or) and radiatum (Ra) but not in the pyramidal layer (P). Staining of the same section with mouse anti-parvalbumin (red) identifies the pyramidal layer.
- Rat hippocampal neurons (16 μg/ml) (Verpelli, C. et al. (2011) J. Biol. Chem. 286, 34839.).
- 1. Dingledine, R. et al. (1999) Pharmacol. Rev. 51, 7.
- 2. Sheng, M. and Lee, S.H. (2001) Cell 105, 825.
- 3. Song, I. and Huganir, R.L. (2002) Trends Neurosci. 25, 578.
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AMPA receptors are members of the glutamate receptor family of ion channels that also include the NMDA and Kainate receptors. The three subfamilies are named after the original synthetic agonists that were identified as selective ligands of each family.
The α-amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) receptor subfamily includes four members AMPA1-AMPA4 that are also known as GluR1-GluR4 respectively.
The functional AMPA channel is believed to be a tetramer, with most neuronal AMPA receptors being actually heterotetramers composed of AMPA1 plus AMPA2 or AMPA2 plus AMPA3, although homotetramers can also be found.
AMPA receptors are permeable to cations Na+, K+ and Ca2+. The Ca2+ permeability is dependent on the presence of AMPA2: whenever this subunit is present, the channel will be impermeable to Ca2+. The Ca2+ permeability of the AMPA2 subunit is determined by the presence of an arginine (R) at a critical site in the pore loop instead of a glutamine (Q) present in the same site in the other AMPA subunits. A post-transcriptional process known as RNA editing determines the presence of this R. Since most AMPA2 subunits in the adult brain have undergone RNA editing and most AMPA receptors contain the AMPA2 subunit, most native AMPA receptors will be impermeable to Ca2+.
Gating of AMPA receptors by glutamate is extremely fast and therefore the AMPA receptors mediate most excitatory (depolarizing) currents in the brain during basal neuronal activity. The depolarization caused by the activation of post-synaptic AMPA receptors is necessary for the activation of NMDA receptors that will open only in the presence of both glutamate and a depolarized membrane.
Synaptic strength, defined as the level of post-synaptic depolarization, can be long term (hence the term long term potentiation, LTP) and therefore induce changes in signaling and protein synthesis in the activated neuron. These changes are associated with memory formation and learning.
Changes in synaptic strength are thought to involve rapid movement of the AMPA receptors in and out of the synapses and a great deal of effort has focused in understanding the mechanisms that govern AMPA receptor trafficking.
Anti-GluR1 (GluA1) (extracellular) Antibody (#AGC-004) is a highly specific antibody directed against an extracellular epitope of the rat ionotropic glutamate receptor 1. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, and live cell imaging applications. It has been designed to recognize GluR1 from human, mouse, and rat samples.
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（1）确定抗体的名字，注意中英文名字、别名、亚型等信息。
（2）确定您的实验类型，Elisa，WB，IHC，ICC，或者是FACS。每种抗体说明书都会列出抗体经验证过适用的实验类型，如果抗体说明书没有提及的应用类型，并不意味着该抗体不适用于此种分析应用类型，而仅是说明尚未经过此种实验验证。
（3）确定实验样本的种属，Human，Mouse，Rat等。我们的抗体说明书都列出该抗体验证过的适用物种，可根据说明书列出已验证的种属来选择适合你实验的抗体。
（4）样本抗原蛋白的结构性质。了解样本抗原蛋白的结构性质有助于选择最合适的抗体，待测样本蛋白的结构域和样本在提取和处理过程中是否会变性，蛋白空间构象的改变，会影响抗体的免疫亲和反应。
（5）单多克隆抗体的选择。一般单克隆抗体特异性强，但亲和力相对小，检测抗原灵敏度相对就低；而多克隆抗体特异性稍弱，但抗体的亲和力强，灵敏度高，但易出现非特异性染色（可以通过封闭等避免）。
2. 二抗选择
（1）种属来源。主要根据一抗种属来源而决定二抗来源，如一抗是小鼠来源，那二抗就买抗小鼠的即可（羊、兔等均可）。
（2）标记物的选择。有HRP、Biotin、荧光素等标记物。免疫组化，WB二抗主要选择HRP，Biotin标记的二抗，而免疫荧光染色可按实验需要选择不同荧光素标记的二抗，如FITC、Cy3、PE等。
2 一抗是属于哪个类或亚类
二抗需与一抗的类别或亚类相匹配。这通常是针对单克隆抗体而言。多克隆抗体主要是IgG类免疫球蛋白，因此相应的二抗就是抗IgG抗体.
单克隆抗体的类别及亚类通常会在产品列表中列出，如果你的一抗是小鼠IgM，那么相应的二抗就应当是抗小鼠IgM，或是抗小鼠IgG抗体。
如果单克隆一抗是小鼠IgG的某一亚类（IgG1，IgG2a，IgG2b，IgG3），那么几乎所有的抗小鼠IgG都可以与之结合，或者你也可以选择专门针对这一亚类的二抗，例如，如果你的一抗是小鼠IgG1，那么你可以选择抗IgG1的二抗，此种抗体在双标记实验中尤其适合。
如果你不知道一抗是哪一类别或亚类，那么抗小鼠IgG是一个不错的选择，因为此种抗体可以识别大多数类型的IgG免疫球蛋白。
